The goal of this Phase I SBIR proposal is to develop a rapid, low-cost, multiplex immunoassay for the detectionof common respiratory tract pathogens, including influenza A (Flu A), influenza B (Flu B), respiratory syncytialvirus (RSV), and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in less than 15 minutes. Thedevelopment of this antigen-based test will accelerate the rapid diagnosis of SARS-CoV-2 infection, along withinfluenza A / influenza B and RSV. Influenza A / influenza B and RSV are the other common causative agents of respiratory tract infections otherthan the ongoing SARS-CoV-2 pandemic. Analyses from the CDC have suggested that there could be 10 timesmore Coronavirus disease 2019 (COVID-19) positive cases than reported cases. 2 This is primarily due to theslow screening process as it mainly depends on the RT-PCR based testing systems. The rapid identification ofSARS-CoV-2 is critical to stop the spread of the infection as it is highly contagious. Apart from SARS-CoV-2, influenza A / influenza B and RSV cause a considerable amount of infections and severe illness. Moreover, influenza A / influenza B and RSVinfections show similar symptoms to that of SARS-CoV-2 infection. Differentiating SARS-CoV-2 infections withFlu or RSV is also much needed as Flu and RSV infections have better treatment options if they are diagnosedearly. Rapid tests would provide a better opportunity to choose the right treatment strategy and stop the spreadof the illness. We propose to develop a rapid, multiplex test for the detection of influenza A / influenza B, RSV, and SARS-CoV-2without any complex instrumentation. Our rapid test uses immobilized antibodies on the membrane for thecapturing and simultaneous qualitative detection of specific antigens for four common respiratory tractpathogens. We will use our innovatively designed sequential injection syringe (SIS) to pre-fill the reagents anddisperse them sequentially into the flow-through assay cassette. The components such as the 2nd antibodyconjugated with biotin, wash buffers, neutravidin-HRP, and tetramethylbenzidine (TMB) with stable hydrogenperoxide will be sequentially dispersed into the assay cassette. After completing the assay, the colored spotsthat develop can be viewed without any special instrumentation and can be interpreted easily. If needed, a readercan be used to further increase the sensitivity and ability to record and send test result to the physicians. Thismultiplex assay can be performed in less than 15 minutes in various point-of-care (POC) and low resourcesettings without any special training. We will demonstrate that our test can deliver rapid results with the samelevel of performance as singleplex tests for each pathogen using clinical specimens. We will also validate ourassay method at our collaborator's lab at the Medical College of Wisconsin. We plan to achieve 100% specificityand higher sensitivity than current lateral flow assays. If successfully developed, this rapid, multiplex test will besimple to perform and inexpensive enough for all sizes of primary-care physician's offices, nursing homes,pharmacies, and community health clinics to adopt the platform.
Public Health Relevance Statement: Project Narrative
A rapid and minimal instrumentation-based multiplex test for influenza A / influenza B, RSV, and SARS-CoV-2 diagnosis is much
needed at point-of-care and low resource settings. Implementation of our rapid, multiplex test will accelerate the
detection and differentiation of infectious agents in symptomatic and asymptomatic patients. This will enable
timely medical responses to treat the patient and stop the spread of the infection.
Project Terms:
